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Preparation of Highly Magnetizable Spheres for Bio-Separations

Author: CaoZuoZuo
Tutor: WanQianHong
School: Tianjin University
Course: Drug analysis
Keywords: Bio - magnetic separation Solvothermal Magnetic microspheres β-amino acid Immobilized enzyme Lipase Chinese herbal medicines DNA extraction
CLC: R944
Type: Master's thesis
Year: 2009
Downloads: 107
Quote: 1
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Abstract


With the rapid development of science and technology, the field of scientific research and production fields generated huge demand for quick and easy separation. Magnetic separation technology is efficient, fast, non-tarnish to set separated and enriched in one many advantages and are widely used in the immunoassay, nucleic acid extraction, gene sequencing, cell separation, enzyme immobilization, chiral separation of life science research important areas. Bio-magnetic separation technology development trend of miniaturization, parallelization, and automation, the core issue is that prepared a uniform particle size, good dispersion, magnetic and prepared a simple magnetic microspheres, in order to ensure the efficient bioseparation process reproducible and controllable. Based on the above background, the paper aims to explore the high responsiveness magnetic microspheres preparation methods and their application in bioseparation. The research work is divided into three parts: (1) high-response magnetic microspheres and modification of surface functional groups; (2) magnetic microspheres immobilized lipase split β-amino acids; (3) magnetic microspheres extract of Chinese medicine material DNA. Using Fe 3 O 4 magnetic microspheres solvothermal method, a series of studies and into a ball mechanism and control factors. By sol - gel method in magnetic microsphere surface functional group modification, FT-IR, XRD, VSM, TEM, HR-TEM characterizations, the microspheres with small particle size, magnetic, good dispersion, etc., laid the foundation for rapid separation and preparation of simple, conducive to the realization of a large-scale production. The magnetic microspheres are two types of applied research, First, by a covalent bond, introducing space a spacer arm covalently bonded to the metal ion chelate these three different methods, will have the chiral separation of characteristics of the lipase (Pseudomonas cepacia lipase) is fixed to the surface thereof, made of the magnetic microspheres as a carrier of the immobilized enzyme. Split the β-amino acid with the immobilized lipase racemates obtained optically pure isomers. For these three different methods, respectively, explored the optimum conditions of immobilization, and the immobilized enzyme prepared by the three methods, the basic enzymatic properties and splitting performance comparison. Groping through enzyme immobilization methods and conditions of generalized immobilized proteins and cells has important reference value. Carry out the modernization of traditional Chinese medicine, Chinese herbal medicines quality control is facing the challenge of a higher level, based on the authenticity of the pros and cons of identification of the molecular biology of nucleic acid identification of Chinese herbal medicines has important guiding significance. The complex composition of Chinese herbal medicine processing leads to degradation of nucleic acid, thus establishing the nucleic acid extraction method of Chinese herbal medicines, meet purity, concentration and integrity requirements to obtain a nucleic acid template, is related to the success of molecular biology identification key. In this experiment, the magnetic microspheres as the adsorbent from Chinese herbal extract DNA, improved extraction method, which provides a new method for Chinese herbal medicines authenticity of the pros and cons identify. Were selected to the roots, stems, leaves, flowers, seeds, fruits, herbal medicines, the validation of the method, showed that this method can effectively extract the DNA of Chinese herbal medicines, and to provide the conditions for subsequent molecular biology experiments.

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